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At least 595 records · Page 33Linked to original sources
PubMed1994

On-line enzymatic amplification by substrate cycling in a dual bioreactor with rotation and amperometric detection.

The amplification approach centered on the cycling of two reversibly interconvertible chemical species sequentially participating in two different enzyme-catalyzed reactions (enzymatic amplification by substrate cycling) has been implemented on-line into a continuous-flow/stopped-flow/continuous-flow operation. The implementation is illustrated with the determination of L-lactate in a dual enzyme reactor containing immobilized lactate oxidase (LOD, EC 1.1.3.x) to catalyze the oxidation of L-lactate by dissolved oxygen. The immobilized LOD was affixed to a rotating disk in the lower part of the flow-through cell. Immobilized lactate dehydrogenase (EC 1.1.1.27), affixed to the top part of the cell regenerates L-lactate with the mediation of beta-NADH as the hydrogen donor. The substrate cycling permits the generation of H2O2 beyond the stoichiometric limitation, and this is detected at a stationary Pt-ring electrode located at the bottom part of the cell. The stationary Pt-ring electrode is positioned concentrically to the rotating disk containing the immobilized LOD. The resulting amplified response permits, in a simple manner, achievement of detection limits as low as 0.3 fmol.liter-1 and allows the processing of 30 samples per hour.

Animals↗
PubMed1975

Lipoic acid content of Escherichia coli and other microorganisms.

A mutant strain of Escherichia coli K-12 requiring lipoic acid. W1485lip 2 (ATCC 25645), was used to develop a turbidimetric assay for lipoic acid and a polarographic assay based on the oxidation of pyruvate by suspensions of lipoic acid-deficient organisms. The turbidmetric assay was more sensitive with a working range equivalent to 0.2-2.0 ng of DL-alpha-lipoic acid compared with 5-50 ng for the polarographic method. The mutant responded equally to racemic mixtures of alpha-lipoic acid, beta-lipoic acid and dihydrolipoic acid but gave little response to lipoamide, and other derivatives without prior hydrolysis; 8-methyllipoic acid was a competitive inhibitor of the response to lipoic acid. A high specificity of the mutant for the natural steroisomer was indicated by the fact that (+)-alpha-lipoic acid had twice the activity of the racemic mixture. Escherichia coli K12 contained less than 0.05 ng of free (+)-alpha-lipoic acid per mg dry weight but, depending on the growth substrate, the equivalent of between 13 and 47 ng of (+)-alpha-lipoic acid per mg dry weight after acid extraction. There was a strong correlation between the lipoic acid content and the sum of the specific activities for the pyruvate and alpha-ketoglutarate dehydrogenase complexes. Experiments with washed suspensions of Escherichia coli showed only small increases in lipoic acid content (18%) when incubated with pyruvate, cysteine and methionine. When supplied with exogenous lipoic acid the mutant, W1485lip2, accumulated very little more than was demanded by its metabolism. The lipoic acid contents of several organisms were measured and correlated with their metabolism.

Bacillus megaterium↗
PubMed1994

The potential of lactic acid bacteria for the production of safe and wholesome food.

By tradition lactic acid bacteria (LAB) are involved in the production of fermented foods. These constitute one quarter of our diet and are characterized by a safe history, certain beneficial health effects, and an extended shelf life when compared with raw materials. The various fermenting substrates are habitats for specific LAB that differ in their metabolic potential. The health effects exerted by LAB are the following: 1. Production of lactic acid and minor amounts of acetic and formic acid. These cause: a drop in pH and thereby growth inhibition of food spoiling or poisoning bacteria; killing of certain pathogens; detoxification by degradation of noxious compounds of plant origin (usually in combination with plant-derived enzymatic activities). 2. Production of antimicrobial compounds (e.g. bacteriocins, H2O2, fatty acids). 3. Probiotic effects as live organisms in food. The wholesomeness of LAB can also be extended to fields outside human nutrition, as they may act as probiotics in animal production or as plant protectives in agriculture and thus contribute to healthy raw materials for food production. Modern concepts or perspectives of the application of LAB include the following: 1. Selection of the best adapted and safely performing LAB strains. 2. Selection of strains with probiotic effects. 3. Selection of strains with health-promoting effects (e.g. production of vitamins or essential amino acids, anti-tumour activity). 4. Selection of strains with food protective activities (inhibiting spoilage or food pathogens). These strains can be added to food or used as starters in food fermentations. They may be found as wild-type organisms or can be obtained by genetic engineering.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed2001

Luminescent method for the detection of antibacterial activities.

A new rapid and sensitive method for the detection of antibacterial activities was based on luminescent indicator strains. Listeria innocua 8811 and Enterococcus faecalis 32 were transformed with plasmid carrying bacterial luciferase genes. Subsequent strains became capable to emit light during the exponential growth phase. The addition of bacteriocin containing culture supernatants to such cultures induced a drop of their light emission which was correlated to the combined antibacterial activity of acid stress and bacteriocin. The detection of antagonistic activity is independent of its mode of action, i.e. bactericidal or bacteriostatic. This method allowed to directly visualize the antagonistic activity of bacteriocin producer strains toward target strains in coculture experiments. However, a control co-culture with non-producing bacteriocin mutant was necessary in order to distinguish between nutrients competition and bacteriocin activity. Finally, five class IIa bacteriocins were purified from culture supernatants of eight strains detected in 3 days from a 120 lactic acid bacteria collection.

Bacteriocins↗